Stimulation of in vitro root and shoot growth of potato by increasing sucrose concentration in the presence of fluridone,an inhibitor of abscisic acid synthesis

Fluridone, an inhibitor of abscisic acid (ABA) biosynthesis, strongly stimulated rooting of nodal stem segments of potato (Solanum tuberosum L.) cultivar Arran Banner cultured in darkness on tuberisation medium. Inclusion of 10^-6 M ABA in the culture medium prevented this rooting response, indicating that root proliferation in the presence of fluridone could be due to inhibition of ABA synthesis. The rooting response to fluridone (increased total root number and root fresh weight) was obtained only at high sucrose concentrations (0.175 and 0.234 M) and was demonstrated with two potato cultivars and two culture media; one which favoured tuberisation and one which did not. Shoot numbers were also increased, but to a lesser extent than root numbers, and total fresh weight of plant material per culture was greatly increased by inclusion of both fluridone (10^-6 or 10^-5 M) and 0.234 M sucrose in the culture medium. The role of sucrose was not simply osmotic because when the osmolarity of fluridone medium was increased using mixtures of mannitol and sucrose, no root proliferation occurred unless sucrose predominated in the mixture.     

A rapid method for preparing high quality alizarin stained skeletons of adult mice

A simple three-day technique is described for preparing completely cleared and high quality alizarin stained total skeletons of adult mice. Unfixed specimens are partially macerated during staining. Older specimens are heated for 15 min in 1% KOH. A heated solution of benzyl and ethyl alcohol, glycerin, and water is used for final clearing and hardening. This procedure requires about 10 min work per specimen and greatly simplifies preparation of stained and cleared skeletons of adult mice. Another technique, giving slightly better preparations, but requiring 11-14 days, is also described.     

Betrayal: radio-tagged Burmese pythons reveal locations of conspecifics in Everglades National Park

The “Judas” technique is based on the idea that a radio-tagged individual can be used to “betray” conspecifics during the course of its routine social behavior. The Burmese python (Python bivittatus) is an invasive constrictor in southern Florida, and few methods are available for its control. Pythons are normally solitary, but from December–April in southern Florida, they form breeding aggregations containing up to 8 individuals, providing an opportunity to apply the technique. We radio-tracked 25 individual adult pythons of both sexes during the breeding season from 2007–2012. Our goals were to (1) characterize python movements and determine habitat selection for betrayal events, (2) quantify betrayal rates of Judas pythons, and (3) compare the efficacy of this tool with current tools for capturing pythons, both in terms of cost per python removed (CPP) and catch per unit effort (CPUE). In a total of 33 python-seasons, we had 8 betrayal events (24 %) in which a Judas python led us to new pythons. Betrayal events occurred more frequently in lowland forest (including tree islands) than would be expected by chance alone. These 8 events resulted in the capture of 14 new individuals (1–4 new pythons per event). Our effort comparison shows that while the Judas technique is more costly than road cruising surveys per python removed, the Judas technique yields more large, reproductive females and is effective at a time of year that road cruising is not, making it a potential complement to the status quo removal effort.     

Sample size determination in clinical proteomic profiling experiments using mass spectrometry for class comparison

Mass spectrometric profiling approaches such as MALDI‐TOF and SELDI‐TOF are increasingly being used in disease marker discovery, particularly in the lower molecular weight proteome. However, little consideration has been given to the issue of sample size in experimental design. The aim of this study was to develop a protocol for the use of sample size calculations in proteomic profiling studies using MS. These sample size calculations can be based on a simple linear mixed model which allows the inclusion of estimates of biological and technical variation inherent in the experiment. The use of a pilot experiment to estimate these components of variance is investigated and is shown to work well when compared with larger studies. Examination of data from a number of studies using different sample types and different chromatographic surfaces shows the need for sample‐ and preparation‐specific sample size calculations.     

Suspension trapping (STrap) sample preparation method for bottom‐up proteomics analysis

Despite recent developments in bottom‐up proteomics, the need still exists in a fast, uncomplicated, and robust method for comprehensive sample processing especially when applied to low protein amounts. The suspension trapping method combines the advantage of efficient SDS‐based protein extraction with rapid detergent removal, reactor‐type protein digestion, and peptide cleanup. Proteins are solubilized in SDS. The sample is acidified and introduced into the suspension trapping tip incorporating the depth filter and hydrophobic compartments, filled with the neutral pH methanolic solution. The instantly formed fine protein suspension is trapped in the depth filter stack—this crucial step is aimed at separating the particulate matter in space. SDS and other contaminants are removed in the flow‐through, and a protease is introduced. Following the digestion, the peptides are cleaned up using the tip's hydrophobic part. The methodology allows processing of protein loads down to the low microgram/submicrogram levels. The detergent removal takes about 5 min, whereas the tryptic proteolysis of a cellular lysate is complete in as little as 30 min. We have successfully utilized the method for analysis of cellular lysates, enriched membrane preparations, and immunoprecipitates. We expect that due to its robustness and simplicity, the method will become an essential proteomics tool.